Monoclonal Antibody Production|2 Methods in Detail
Monoclonal antibodies are set of antibodies which are target specific against a single source of antigen.
In every living organism, they are produced by white blood cells as a part of immune mechanism.
They are obtained from single set of cells and hence named as monoclonal antibodies.
Monoclonal Antibody Production is taken up in industrial scale as they are extensively used in medical field.
There is a huge demand due to their application in biomedical research, diagnosis, treatment of infectious.
They are also used in treatment of cancer.
Before we go further, we assume that you know what is immunity, antigen and antibody.
Production of monoclonal antibodies is done of by either of two methods.
- In Vitro Method (Using cell lines in glass ware)
- In Vivo method (Using living animals like Mice)
involves following steps
1. First an animal like a mouse, rabbit etc. is immunized with suitable antigen which corresponds to desired monoclonal antibody we are interested in. This is done by injecting antigens which are previously emulsified with some adjuvants like Freund’s adjuvant.
2. This leads to enhanced production of desired antibodies in the mouse body. The immunization is done for few weeks till the antibody concentration in the mouse blood reaches a desired level.
3. After several weeks, the blood test is done to see antibody titre using suitable techniques like ELISA or flow cytometry etc.and are obtained from mouse spleen.
4. Then the obtained monoclonal antibodies are fused with a cancer cell (i.e. myeloma not just tumor) as they are immortal and can divide indefinitely.
5. So the fused cell grow indefinitely to produce huge number of monoclonal antibodies..The bunch of fused cells is called hybridoma.
6. These formed tumor cells are injected into mouse peritoneal cavity or grown in vitro by tissue culture technique.
8. On injection into mouse peritoneal cavity, the cell grow rapidly and release them in the form of liquid into the abdomen which appear as bulged fluid bags (ascites).
9. These monoclonal antibodies from ascites are extracted to harvest the formed antibodies.
Another method is growing these hybridoma cells invitro but is not easy and also is expensive than the mouse method. But sometimes the mouse may experience pain, irritation due to ascites. Then they go for invitro cell culture technique.